Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [2]
- Flow cytometry [1]
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Validation data
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- Product number
- MA5-32779 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Galectin 1 Recombinant Rabbit Monoclonal Antibody (JM13-37)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JM13-37
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Galectin 1 in HL-60 cell (1) and human skeletal muscle tissue (2) lysate using a Galectin 1 Monoclonal antibody (Product # MA5-32779) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Galectin 1 was achieved by transfecting PC-3 with Galectin 1 specific siRNAs (Silencer® select Product # s194592, s8145). Western blot analysis (Fig. a) was performed using whole cell extracts (25ug lysate) from the Galectin 1 knockdown cells (Lane 3), non-targeting scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Galectin 1 Recombinant Rabbit Monoclonal Antibody (JM13-37) (Product # MA5-32779, 1:2500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:8000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Galectin 1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Galectin 1 Recombinant Rabbit Monoclonal Antibody (JM13-37) (Product # MA5-32779) and a 15kDa band corresponding to Galectin 1 was observed in MDA-MB-231, THP-1, PC-3 and Caki-1 but not in MCF7, LNCaP. Whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), MCF7 (Lane 2), THP-1 (Lane 3), LNCaP (Lane 4), PC-3 (Lane 5) and Caki-1 (Lane 6) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1250 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:8000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Increased expression of Galectin 1 is seen in triple negative breast cancer (TNCB) (like MDA-MB-231) for promoting metastasis but not in non-TNBC like MCF7 [10.18632/oncotarget.16208] and in androgen unresponsive PC-3 cells in comparison to androgen responsive LNCaP [10.1155/2013/519436]
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of Galectin 1 in A431 cells using a Galectin 1 Monoclonal antibody (Product # MA5-32779) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Galectin 1 was performed using 70% confluent log phase MDA-MB-231 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Galectin 1 Recombinant Rabbit Monoclonal Antibody (JM13-37) (Product # MA5-32779) at 1:200 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2500 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization of Galectin 1 in MDA-MB-231 cells. Panel e represents MCF7 cells showing negative staining for the same. Panel f represents control MDA-MB-231 cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Galectin 1 of paraffin-embedded rat stomach tissue using a Galectin-1 Monoclonal antibody (Product #MA5-32779). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Galectin 1 of paraffin-embedded Human placenta tissue using a Galectin-1 Monoclonal antibody (Product #MA5-32779). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of Galectin 1 in NIH-3T3 cells using a Galectin 1 Monoclonal Antibody (Product # MA5-32779) at a dilution of 1:100, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black).